Cryopreservation of Dioscorea rotundata poir.: a comparative study with two cryogenic procedures and assessment of true-to-type of regenerants by RAPD analysis.
The aim of this study was to develop a cryopreservation protocol for Dioscorea rotundata with maintenance of genetic stability of regenerated plants after cryopreservation. In vitro shoot tips were cryopreserved using vitrification and encapsulation-dehydration to compare the efficacy of the two methods. Both methods produced high levels of plant regeneration from cryopreserved shoot tips. The regeneration level obtained using vitrification (71%) was not significantly different from that obtained using encapsulation-dehydration (67%). Genetic stability of plants derived from cryopreserved shoot tips was evaluated using RAPD markers. Analysis of 50 cryopreserved-derived and 20 in vitro- maintained (control) plantlets showed that 10 primers produced 77 clear, reproducible bands, with the amplification products being monomorphic for all the plantlets tested. A total of 5,390 bands obtained from this study exhibited no aberration in RAPD banding. Thus, the present study showed that both vitrification and encapsulation-dehydration methods are equally applicable to D. rotundata for cryopreservation. The in vitro plantlets derived from cryopreservation were genetically stable at the molecular level tested.
Mandal
B B
BB
Tissue Culture and Cryopreservation Unit, National Bureau of Plant Genetic Resources, New Delhi, India. mandalbinay@yahoo.co.in
Ahuja-Ghosh
Sangeeta
S
Srivastava
P S
PS
D015925:Cryopreservation / Q000379:methods*; D027742:Dioscorea / Q000235:genetics / Q000254:growth & development*; D018520:Plant Shoots / Q000235:genetics / Q000254:growth & development*; D019105:Random Amplified Polymorphic DNA Technique